Methods, kits and compositions for single primer linear isothermal amplification of nucleic acid sequences
First Claim
1. A kit for amplifying a target RNA or the complement of a target RNA comprising a first composite primer, a second composite primer, and a third composite primer, wherein each composite primer comprises an RNA portion and a 3′
- DNA portion, and wherein the third composite primer is a composite amplification primer and comprises a sequence that is hybridizable to a polynucleotide comprising a complement of the first composite primer.
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Accused Products
Abstract
The invention provides methods for isothermal amplification of RNA. The methods are particularly suitable for amplifying a plurality of RNA species in a sample. The methods employ a composite primer, a second primer and strand displacement to generate multiple copies of DNA products comprising sequences complementary to an RNA sequence of interest. In another aspect, the methods employ a single primer (which is a composite primer) and strand displacement to generate multiple copies of DNA products comprising sequences complementary to an RNA sequence of interest. In some embodiments, a transcription step is included to generate multiple copies of sense RNA of an RNA sequence of interest. The methods are useful for preparation of nucleic acid libraries and substrates for analysis of gene expression of cells in biological samples. The invention also provides compositions and kits for practicing the amplification methods, as well as methods which use the amplification products.
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Citations
26 Claims
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1. A kit for amplifying a target RNA or the complement of a target RNA comprising a first composite primer, a second composite primer, and a third composite primer, wherein each composite primer comprises an RNA portion and a 3′
- DNA portion, and wherein the third composite primer is a composite amplification primer and comprises a sequence that is hybridizable to a polynucleotide comprising a complement of the first composite primer.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
Specification